Tall fescue regeneration plant obtaining method through tissue culture
A technology for regeneration of plants and tissue culture, applied in the biological field, can solve the problems of significant differences in regeneration ability and difficult regeneration of varieties
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Embodiment 1
[0026] The American tall fescue cultivar 'surpro' was used as the test material, which was purchased from the Lawn Research Institute of China Agricultural University. Choose 150 plump seeds, soak them in 75% alcohol for 3 minutes after removing their shells, then disinfect them with 0.1% mercuric chloride for 10 minutes, and then wash them thoroughly with sterile distilled water 5 times. Disinfected seeds were inoculated into the callus induction medium MSM containing 5mg / L, 7mg / L, 9mg / L 2,4-D and three treatments of 0.025mg / L KT (50 seeds for each treatment). Wounded tissue induction culture. After 2 weeks, the calli induced by each treatment were transferred to the callus maintenance medium containing 4.5mg / L2, 4-D and 0.2mg / L KT in MSM for proliferation. After 3 weeks of callus proliferation and culture, they were divided into mung bean sizes and transferred to MSM medium containing 2mg / L 2,4-D and 0.2mg / L KT (hydrolyzed casein content was 1mg / L) for somatic embryogenesis...
Embodiment 2
[0028]The American tall fescue cultivar 'Coronado' was purchased from the Lawn Research Institute of China Agricultural University. Choose 150 plump seeds, soak them in 75% alcohol for 3 minutes after removing their shells, and then disinfect them with 0.1% mercuric chloride for 10 minutes, and then wash them thoroughly with sterile distilled water 5 times. The sterilized seeds were inoculated into the callus induction medium MSM containing 6mg / L, 8mg / L, 10mg / L 2, 4-D and 0.025mg / L KT under three treatments (50 seeds for each treatment). Wounded tissue induction culture. After 2 weeks, the callus formed under each treatment was transferred to the callus maintenance medium containing 4.5mg / L2, 4-D and 0.2mg / L KT (hydrolyzed casein content was 2mg / L) Proliferate. After callus proliferation and culture for 4 weeks, the callus with somatic embryos was inoculated on the differentiation medium MSH+2mg / L KT+2% sucrose (the content of hydrolyzed casein was 2mg / L) to differentiate in...
Embodiment 3
[0030] The European cultivar 'Summer Lawn' was used as the test material, which was purchased from the Lawn Research Institute of China Agricultural University. Choose 150 plump seeds, soak them in 75% alcohol for 3 minutes after removing their shells, then disinfect them with 0.1% mercuric chloride for 10 minutes, and then wash them thoroughly with sterile distilled water 5 times. Disinfected seeds were inoculated into the callus induction medium MSM containing 5mg / L, 7mg / L, 9mg / L 2,4-D and three treatments of 0.025mg / L KT (50 seeds for each treatment). Wounded tissue induction culture. After 2 weeks, the callus formed under each treatment was transferred to the callus maintenance medium containing 4.5mg / L2, 4-D and 0.2mg / L KT (hydrolyzed casein content was 1mg / L) Proliferate. After 3 weeks of callus proliferation and culture, they were transferred to MSM medium containing 2mg / L 2,4-D and 0.1mg / L KT (hydrolyzed casein content was 1mg / L) for induction culture of somatic embr...
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